Many diverse diseases, including cataract, some types of glaucoma, Alzheimer disease, diabetes and cancer to name only a few, are now known to share the common feature of aggregated, misfolded or modified protein deposits. The pathological hallmark in this group of diseases is protein aggregation and deposition in specific cells, tissues or organs. The pathological similarities indicate that common principles that govern protein interactions underlie protein misfolding degenerative diseases. Cataract is a classic example of a protein misfolding and aggregation disease. Cataract, a major cause of blindness in the world, develops as a result of age-related modifications and aggregation of the lens proteins. ?-crystallin accounts for nearly 40% of the adult lens proteins but its structure-function is yet to be fully understood. The chaperone-like activity of ?-crystallin is believed to play a central role in maintaining lens transparency. During aging, lens crystallins undergo truncation and these modifications correlate with lens crystallin aggregations responsible for light scattering. It is well established that the addition of ?-crystallin to aggregating proteins stops a further increase in aggregation and light scattering. We have demonstrated that specific sequences in ?-crystallin subunits suppress aggregation of denaturing proteins. However, the full potential of ?-crystallin-derived peptides (mini-chaperones) is yet to be realized. To enhance our understanding of the role of non-native interactions in cataract formation, we propose the following specific aims: Aim 1. Identify the sequences involved in abnormal interactions of ?A-crystallin subunits that lead to protein aggregation in human cataract-causing ?A-crystallin mutants (?AR49C, ?AF71V, ?AG98R and ?AR116H) using novel deuterated cross linkers, biotynylated reagents and peptide arrays. Aim 2. Determine the mechanism and efficacy of ?A-crystallin-derived mini-chaperone in preventing the aggregation of metastable ?A- crystallins (mutants and truncated crystallins) and restoring chaperone activity. The specific aims will be accomplished using novel cross-linkers and mass spectrometric methods. We will also use site-directed mutagenesis and cell culture expression and analysis systems to confirm the cross linking data. These innovative studies will give us new insights into potential interventions for protein misfolding diseases, not only of the eye but also of other parts of the body.